rabbit anti human transferrin receptor 1 antibody  (Cell Signaling Technology Inc)

Journal: Neural Development

Article Title: Analysis of Lrrn1 expression and its relationship to neuromeric boundaries during chick neural development

doi: 10.1186/1749-8104-2-22

Figure Lengend Snippet: Endosomal distribution of Lrrn1. Immunocytochemical analysis of the endosomal uptake of FLAG-Lrrn1 in HeLa cells. (a, d, g, j) Plasma membrane surface-exposed FLAG-Lrrn1 protein detected with an anti-FLAG antibody. (b, c) Colocalisation with total cellular pool of Lrrn1 detected with the GST-IC antiserum. (e, f) Colocalisation with EEA1 (arrowheads in inset in (f)) shows that a significant pool of Lrrn1 cycles through the early endosomes. (h, i) Little overlap with transferrin receptor immunostaining suggests that Lrrn1 does not partition within recycling endosomes. (k-m) A very small proportion of endosomes were found to co-label with FLAG-Lrrn1/EGF ((m), arrowhead in inset). Scale bars = 200 μm except inset in (f) = 50μm.

Article Snippet: The antibody against EEA1 (rabbit polyclonal antiserum 1:1,500) was a kind gift of Dr Mario Zerial (EMBL, Heidelberg, Germany) and the anti-human transferrin receptor (rabbit polyclonal, 1:250) was from Chemicon.

Techniques: Immunostaining

Journal: Hepatology (Baltimore, Md.)

Article Title: Human macrophage ferroportin biology and the basis for the ferroportin disease

doi: 10.1002/hep.29007

Figure Lengend Snippet: WT and mutant FPN1 cycles in the early endosome/plasma membrane compartment in human macrophages. (A) Coimmunofluorescence staining of FPN1 (green) with markers (red) for early endosomes, EE (EEA‐1), lysosomes (LAMP‐1), plasma membrane, PM (Na,K‐ATPase), Golgi (golgin‐97), and endoplasmatic reticulum, ER (calnexin), shows that FPN1 colocalizes with early endosomes and lysosomes (merge panels, yellow) in p.A77D, p.G80S, and pVal162del macrophages, whereas it is almost exclusively localized in the EE compartment, in p.A69T and donor macrophages. (B) Coimmunostaining of FPN1 (green) with TfR1 (red; a marker for the early endocytic recycling compartment) shows that FPN1 colocalizes with TfR1 in donor as well as in p.G80S and p.A77D macrophages. (C) Treatment of macrophages from donors and p.A77D patients with dynasore to block dynamin‐dependent endocytosis leads to accumulation of both FPN1 and TFR1 in the PM compartment.

Article Snippet: The primary antibodies used were: mouse antihuman CD14 (1:20; Chemicon International, Temecula, CA); mouse anti–sodium‐potassium pump (Na,K‐ATPase [plasma membrane marker]; 1:500, Abcam, Cambridge, UK); rabbit anticalnexin (1:200 [endoplasmic reticulum marker]; Sigma‐Aldrich); rabbit anti‐golgin‐97 (1:100 [Golgi marker]; Molecular Probes, Leiden, The Netherlands )(Golgi marker); rabbit anti–early endosome antigen 1 (EEA‐1; 1:200; Abcam, Cambridge, UK), as an early endosomes marker; rat anti–lysosomal‐associated membrane protein 1 (LAMP‐1 [lysosome marker]; 1:50; Fitzgerald Industries International, Concord, MA); rabbit anti‐FPN1 (1:50 PBS; Alpha Diagnostic International, San Antonio, TX, USA); rabbit anti–human transferrin receptor 1 (TfR1; 1:50; Santa Cruz Biotechnology, Santa Cruz, CA); and rabbit anti‐Hepc (1:50 PBS; Alpha Diagnostic International).

Techniques: Mutagenesis, Staining, Marker, Blocking Assay